Browsing by Subject "paksusuolisyöpä"

Background and aim: Colorectal cancers (CRC) are the third most common type of cancer in men and the second most common in women worldwide. The risk for CRC is influenced by lifestyle factors, especially diet. The effect of diet on CRC risk is thought to be mediated by the liquid phase of the feces called fecal water (FW); the components in food or produced from food by the microbiome must be in soluble form to have an effect on the colon epithelial cells, especially on the proliferating cells at the bottom of the colonic crypts, and consequently on CRC risk. In this study, we examined how viability of cells from human colon cancer cell line HCA-7 was affected, when the cells were exposed to FW extracted from fecal samples from study participants eating diets with emphasis on either animal or plant protein sources. Additionally we tested, whether body mass index (BMI), pH of the FW, amount of N-nitroso compounds in feces (NOC), or intake of fiber, protein, calcium or energy correlate with viability. Materials and methods: 146 healthy voluteers (age 20–69, BMI 18,5–35) were randomly assigned to three intervention diet groups for 12 weeks. Protein content of the diets (target 17 E%) came from animal and plant sources in proportion of 70/30 % (ANIMAL-group, n=46), 50/50 % (50/50-group, n=44) and 30/70 % (PLANT-group, n=44). Food consumption was assessed using 4-day food records kept by the participants before and at the end of the intervention period. Fecal samples were collected from the same time periods. Fecal samples were diluted with water in ratio of 1:1 and homogenized, homogenate was ultracentrifuged, pH of the supernatant i.e. FW was measured and finally the FW was filtered sterile using a 0,22 μm filter. HCA-7-cells were first incubated roughly 24 h on 96-well cell culture plate in growth medium so that the cells were attached to the plate. Then three parallel FW samples in three different dilution (10 vol-%, 20 vol-% and 30 vol-%) from each study participant were added on to the plate, and the plate was further incubated exactly 24 h. After that, the amount of living cells was measured colorimetrically using WST-8-reagent. Viability (%) was determined by calculating the proportion of living cells in sample containing wells compared to the living cells in the control wells. Statistical significance of the differences of viability between the diet groups was tested by analysis of covariance (ANCOVA) using the viability at the baseline as covariate. Correlations were analyzed using Pearson’s correlation test. Results: FW samples were obtained from134 study participants. There was a statistically significant difference in viability between the groups’ means in samples with FW content of 30 vol-% (ANCOVA p=0,005). Viability (%) was smallest in the PLANT-group (co-variate adjusted mean 30,0 ± 1,9), and in the reciprocal comparison it differed from both the ANIMAL-group (37,8 ± 1,8; p = 0,011, Bonferroni correction) and the 50/50-group (37,1 ± 1,8; p = 0,021). At the end of the study, there were statistically significant correlation (p < 0,05) between viability and BMI in all of the FW contents (10 vol-%: Pearson correlation coefficient R = 0,210; 20 vol-%: R = 0,366; 30 vol-%: R = 0,319), pH of the FW in the FW contents of 20 vol-% (R = 0,204) and 30 vol-% (R = 0,249). The intake of protein (g/d) correlated in contents of 10 vol-% (R = 0,199) and 30 vol-% (R = 0,181). Also, the intake of protein in proportion to energy intake (E%) correlated in contents of 10 vol-% (R = 0,179) and 30 vol-% (R = 0,236). The intake of fiber, calcium and the amount of NOC:s in feces did not correlate with viability. Conclusions: In this study, the FW from the PLANT group reduced the viability of HCA-7 cells in vitro compared to the FW of the 50/50 ja the ANIMAL groups. This could indicate that increasing the consumption of plant-based protein sources and decreasing that of animal proteins may decrease the risk of CRC. To strengthen the current results, the exposure experiments should be replicated also using different colon cancer cell lines and a normal-like control cell line.

Paksusuolisyöpä on yksi yleisimmistä syöpätyypeistä, ja sen riskin tiedetään olevan yhteydessä joihinkin ravintotekijöihin. Esimerkiksi marjojen, hedelmien ja vihannesten runsas käyttö voi suojata paksusuolisyövältä. Marjojen, niiden sisältämien polyfenoleiden sekä marjoista valmistettujen uutteiden on aiemmin havaittu hillitsevän syöpäsolujen kasvua in vitro -solumalleissa. Kotimaisesta hillasta valmistettua uutetta on kuitenkin tutkittu verrattain vähän. Tämän tutkimuksen tavoitteena oli tutkia hillauutteen vaikutusta paksusuolisyövän solujen kasvuun, morfologiaan ja tiettyihin syöpäsensitiivisiin proteiineihin kaksi- ja kolmiulotteisessa soluviljelmässä. Tarkastelun kohteena oli solujen viabiliteetti sekä neljä syövän kehittymiseen yhteydessä olevaa proteiinia: beta-kateniini, E-kadheriini, aktiivinen kaspaasi 3 sekä Ki-67. Koska 3D-solumallia ei vielä ole hyödynnetty HY:n ravitsemustieteen osastolla ravitsemustutkimuksessa, tavoitteena oli myös testata käytössä olleen menetelmän toimivuutta ravinnon yhdisteiden ja syövän yhteyden tutkimisessa. Tutkimuksessa käytettiin ihmisen paksusuolisyövän HCA7-linjan soluja. Altisteena käytetty hillauute oli HY:n elintarvikekemian laboratorion valmistama ja sisälsi 218 µl/ml ellagiinihappoa. Kontrolliliuoksena käytettiin hillauutteen liuotinta dimetyylisulfoksidia (DMSO). Lisäksi kokeet kontrolloitiin näytteillä, joita ei altistettu kummallekaan liuokselle vaan kasvatettiin vastaava aika tavanomaisessa kasvatusliuoksessa. Hillauutteen vaikutusta solujen viabiliteettiin mitattiin MTT-reagenssiin pohjautuvalla solulaskentamenetelmällä. Solujen morfologiassa tapahtuviin syöpäsensitiivisiin proteiineihin liittyviä muutoksia määritettiin vasta-ainespesifisen immunofluoresenssivärjäyksen ja laserkonfokaalimikroskopian avulla. Hillauute ja sen kontrolliliuos DMSO hillitsivät HCA7-solujen kasvua kaksi- ja kolmiulotteisissa malleissa verrattuna soluihin, jotka kasvoivat tavanomaisessa kasvatusliuoksessa. Solujen viabiliteetti oli 30–60 % verrattuna kontrollisoluihin, kun hillauutetta ja DMSO:ta lisättiin elatusaineeseen 75 µl/ml. Viabiliteetissä oli annos-vastesuhde, joka ei ollut lineaarinen. Hillauute lisäsi apoptoosin lisääntymistä ennustavan aktiivisen kaspaasi-3:n sekä soluliitoksissa esiintyvän beta-kateniinin määrää kolmiulotteisessa soluviljelmässä verrattuna DMSO- ja kontrollisoluihin. Muiden tutkittujen syöpäsensitiivisten proteiinien määrissä ei ollut eroja kontrolli- ja uuteviljelmien välillä. Sekä DMSO että hillauute hillitsevät HCA7-solujen kasvua suurina pitoisuuksina. Hillauute vaikuttaisi kuitenkin hillitsevän solujen kasvua lisäämällä apoptoosia, mikä ilmenee kaspaasi-3:n määrän lisääntymisenä hillauutteelle altistuneissa soluissa. Tämän perusteella DMSO:ssa kasvaneet solut kuolivat enemmän nekroosin välityksellä. Hillauutteessa kasvaneissa soluissa beta-kateniini lokalisoitui enemmän solukalvoille, mikä viittaa niiden välisten soluliitosten olleen vahvempia kuin kontrolli- tai DMSO-soluissa.

Backround and aim of the sudy: Berries and berry polyphenols inhibit the growth of cancerous tumors in vivo and cancer cells in vitro. In the digestive system polyphenols are converted to various metabolites, such as phenolic acids, which may partially count for polyphenol bioactivity. These and other components in the liquid fraction of feces, i.e. the fecal water, are in contact with the proliferating cells residing at the bottom of colonic crypts, and may therefore affect colon cancer risk. The aim of this study was to investigate whether berry consumption changes the effect of fecal water on colon cancer cell viability in vitro, when the amount of dietary red meat is controlled. Also of interest was to investigate the possible correlation between fecal water phenolic acids and the viability of fecal water-exposed cells. Materials and methods: Karnimari was a four-week randomized intervention study with two study groups; one receiving 200 g berries and 150 g pork products daily, and the other group receiving pork products only. Fecal samples were collected before and after intervention and fecal waters were extracted. Polyphenol metabolites from end-point fecal waters were analyzed using UHPLC-DAD-FLD-method. Human colon cancer cells (Caco-2 ja HCA-7) and monkey fibroblasts (CV1-P) were exposed to diluted fecal waters (10 % / 20 % / 30 % fecal water) for 24 h, and cell viability was measured with a method based on cellular dehydrogenase activity. The difference in cell viability between groups after intervention was tested with covariance analysis (ANCOVA). The difference in fecal water concentration of phenolic acids between groups after intervention and the difference in cell viability between groups before intervention were tested with Mann-Whitney U-test. The correlations between fecal water phenolic acids and cell viability were tested with Spearman’s correlation. Results: After intervention fecal waters from the berry group reduced cell viability more than fecal waters from the control group. The difference was significant in all three cell lines when fecal water concentration in cell medium was 30 % (HCA-7 p<0.001; Caco-2 p=0.032; CV1-P p=0.009), and in cells HCA-7 (p=0.007) and CV1-P (p=0.003) also when fecal water concentration was 20 %. The concentrations of fecal water protocathequic acid (p=0.027) and p-coumaric acid (p=0.003) were greater in the berry group than in the control group. p-Coumaric acid concentration was significantly correlated with cell viability in all cell lines when fecal water concentration in cell medium was 20 % or 30 %, regardless of the relatively small amounts of p-coumaric acid detected. Conclusions: Fecal waters from the berry group reduced the viability of both colon cancer cell lines and fibroblasts more than fecal waters from the control group. Eating berries led to greater amounts of protocathequic acid and p-coumaric acid in fecal water. p-Coumaric acid may reduce cell viability or be connected to another affective component in fecal water. The biological mechanisms behind the cell viability reducing ability of fecal water from berry eating subjects should be further investigated.